重组Anti-MEKK3抗体[ep600y] (ab40756)
![Immunoprecipitation - Anti-MEKK3 antibody [ep600y] (ab40756)](https://www.abcam.cn/ps/products/40/ab40756/Images/ab40756-501437-antimekk3-antibody-ep600y-immunoprecipitation-mouse.jpg "Immunoprecipitation - Anti-MEKK3 antibody [ep600y] (ab40756)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [ep600y] to MEKK3
- Suitable for: ICC/IF, IHC-P, Flow Cyt (Intra), WB, IP
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-MEKK3抗体[ep600y]
参阅全部 MEKK3 一抗 -
描述
兔单克隆抗体[ep600y] to MEKK3 -
宿主
Rabbit -
经测试应用
适用于: ICC/IF, IHC-P, Flow Cyt (Intra), WB, IPmore details -
种属反应性
与反应: Mouse, Human -
免疫原
Synthetic peptide within Human MEKK3 aa 600-700 (C terminal). The exact sequence is proprietary.
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阳性对照
- ICC: HeLa cells. IHC-P: Human ovary carcinoma. Flow Cyt (intra): HeLa and Raji cells. WB: Human MEKK3 full-length recombinant protein with GST-Tag, HeLa, bEnd.3, NIH/3T3 whole cell lysates. IP: bEnd.3 whole cell lysate.
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading... -
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
ep600y -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab40756于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| ICC/IF |
1/100 - 1/250.
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| IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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| Flow Cyt (Intra) |
1/100 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
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| WB |
1/200 - 1/1000. Detects a band of approximately 70 kDa (predicted molecular weight: 71 kDa).
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| IP |
1/20 - 1/30.
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| 说明 |
|---|
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ICC/IF
1/100 - 1/250. |
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IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
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Flow Cyt (Intra)
1/100 - 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
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WB
1/200 - 1/1000. Detects a band of approximately 70 kDa (predicted molecular weight: 71 kDa). |
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IP
1/20 - 1/30. |
靶标
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功能
Component of a protein kinase signal transduction cascade. Mediates activation of the NF-kappa-B, AP1 and DDIT3 transcriptional regulators. -
序列相似性
Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase kinase subfamily.
Contains 1 OPR domain.
Contains 1 protein kinase domain. - Information by UniProt
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数据库链接
- Entrez Gene: 4215 Human
- Entrez Gene: 26406 Mouse
- Omim: 602539 Human
- SwissProt: Q99759 Human
- SwissProt: Q61084 Mouse
- Unigene: 29282 Human
- Unigene: 27041 Mouse
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别名
- M3K3_HUMAN antibody
- MAP/ERK kinase kinase 3 antibody
- MAP3K3 antibody
see all
图片
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Immunoprecipitation - Anti-MEKK3 antibody [ep600y] (ab40756)
71 kDa MEKK3 was immunoprecipitated from 0.35mg of bEnd.3 (mouse brain endothelial cell) whole cell lysate with ab40756 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab40756 at 1/200 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1 (Input): bEnd.3 (mouse brain endothelial cell) whole cell lysate 10 μg.
Lane 2 (+): bEnd.3 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab40756 in bEnd.3 whole cell lysate
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Exposure time: 180 seconds.
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![Western blot - Anti-MEKK3 antibody [ep600y] (ab40756)](https://www.abcam.cn/ps/products/40/ab40756/Images/ab40756-501435-antimekk3-antibody-ep600y-western-blot-mouse.jpg)
Western blot - Anti-MEKK3 antibody [ep600y] (ab40756)All lanes : Anti-MEKK3 antibody [ep600y] (ab40756) at 1/200 dilution
Lane 1 : bEnd.3 (mouse brain endothelial cell) whole cell lysate
Lane 2 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 71 kDa
Observed band size: 71 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Exposure Time: Lane 1: 92 seconds, Lane 2: 180 seconds.
This blot was developed using a high sensitivity ECL substrate.
In lane 2, the lysate was freshly made and used for Western Blotting immediately to minimize protein degradation. -
![Western blot - Anti-MEKK3 antibody [ep600y] (ab40756)](https://www.abcam.cn/ps/products/40/ab40756/Images/ab40756-285573-anti-mekk3-antibody-ep600y-western-blot.jpg)
Western blot - Anti-MEKK3 antibody [ep600y] (ab40756)All lanes : Anti-MEKK3 antibody [ep600y] (ab40756) at 1/1000 dilution
Lane 1 :Recombinant human MEKK2 protein (ab135009)
Lane 2 :Recombinant human MEKK3 protein (ab85297)
Lysates/proteins at 0.015 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 71 kDa
Observed band size: 97 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking and diluting buffer: 5% NFDM/TBST
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![Immunocytochemistry/ Immunofluorescence - Anti-MEKK3 antibody [ep600y] (ab40756)](https://www.abcam.cn/ps/products/40/ab40756/Images/ab40756-53860-anti-mekk3-antibody-ep600y-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-MEKK3 antibody [ep600y] (ab40756)Immunofluorescent staining of HeLa cells using ab40756 diluted 1:100 -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEKK3 antibody [ep600y] (ab40756)](https://www.abcam.cn/ps/products/40/ab40756/Images/ab40756-53861-anti-mekk3-antibody-ep600y-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEKK3 antibody [ep600y] (ab40756)Immunohistochemical analysis of paraffin-embedded human ovary carcinoma using ab40756 diluted 1:100
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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![Flow Cytometry (Intracellular) - Anti-MEKK3 antibody [ep600y] (ab40756)](https://www.abcam.cn/ps/products/40/ab40756/Images/ab40756-6-anti-mekk3-antibody-ep600y-flowcytometry-hela-human.jpg)
Flow Cytometry (Intracellular) - Anti-MEKK3 antibody [ep600y] (ab40756)Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling MEKK3 with purified ab40756 at 1/1000 dilution (1 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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![Flow Cytometry (Intracellular) - Anti-MEKK3 antibody [ep600y] (ab40756)](https://www.abcam.cn/ps/products/40/ab40756/Images/ab40756-7-anti-mekk3-antibody-ep600y-flowcytometry-raji-human.jpg)
Flow Cytometry (Intracellular) - Anti-MEKK3 antibody [ep600y] (ab40756)Flow Cytometry analysis of Raji (Human Burkitt's lymphoma B lymphocyte) cells labeling MEKK3 with purified ab40756 at 1/100 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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![Western blot - Anti-MEKK3 antibody [ep600y] (ab40756)](https://www.abcam.cn/ps/products/40/ab40756/Images/ab40756-53862-anti-mekk3-antibody-ep600y-western-blot.jpg)
Western blot - Anti-MEKK3 antibody [ep600y] (ab40756)Anti-MEKK3 antibody [ep600y] (ab40756) at 1/1000 dilution + HeLa cell lysate at 10 µg
Predicted band size: 71 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted? -
![Anti-MEKK3 antibody [ep600y] (ab40756)](https://www.abcam.cn/ps/products/40/ab40756/Images/ab40756-5-benefits-of-recombinant-antibodies.png)
Anti-MEKK3 antibody [ep600y] (ab40756)
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
文献 (9)
ab40756 被引用在 9 文献中.
- Fang H et al. MiR-132-3p Modulates MEKK3-Dependent NF-?B and p38/JNK Signaling Pathways to Alleviate Spinal Cord Ischemia-Reperfusion Injury by Hindering M1 Polarization of Macrophages. Front Cell Dev Biol 9:570451 (2021). PubMed: 33644040
- Liu P et al. ETV5-mediated upregulation of lncRNA CTBP1-DT as a ceRNA facilitates HGSOC progression by regulating miR-188-5p/MAP3K3 axis. Cell Death Dis 12:1146 (2021). PubMed: 34887384
- Zhang Y et al. Overexpression of MAP3K3 promotes tumour growth through activation of the NF-?B signalling pathway in ovarian carcinoma. Sci Rep 9:8401 (2019). PubMed: 31182739
- Chu W et al. Lovastatin exerts protective effects on endothelial cells via upregulation of PTK2B. Exp Ther Med 12:1741-1749 (2016). PubMed: 27602089
- Lu H et al. The expression and role of MEKK3 in renal clear cell carcinoma. Anat Rec (Hoboken) 298:727-34 (2015). PubMed: 25388155
- Cao XQ et al. MEKK3 and survivin expression in cervical cancer: association with clinicopathological factors and prognosis. Asian Pac J Cancer Prev 15:5271-6 (2014). PubMed: 25040987
- Halaban R et al. PLX4032, a selective BRAF(V600E) kinase inhibitor, activates the ERK pathway and enhances cell migration and proliferation of BRAF melanoma cells. Pigment Cell Melanoma Res 23:190-200 (2010). PubMed: 20149136
- Matitau AE & Scheid MP Phosphorylation of MEKK3 at threonine 294 promotes 14-3-3 association to inhibit nuclear factor kappaB activation. J Biol Chem 283:13261-8 (2008). PubMed: 18308725
- Son MY et al. Expression profiles of protein tyrosine kinase genes in human embryonic stem cells. Reproduction 136:423-32 (2008). PubMed: 18583472